Construction of an Immunized Rabbit Phage Display Library for Selecting High Activity against Bacillus thuringiensis Cryl F Toxin Single-Chain Antibodies

文献类型： 外文期刊

第一作者： Xu, Chongxin

作者： Xu, Chongxin;Zhang, Cunzheng;Zhong, Jianfeng;Zhang, Xiao;Liu, Yuan;Liu, Xianjin;Hu, Hui;Luo, Shimin;Liu, Xiaoqin

作者机构：

关键词： phage display library;rabbit scFv;Bacillus thuringiensis;CrylF toxin;ELISA

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.279； 五年影响因子：5.269 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： In the present study, a Cry1F-immunized rabbit phage display library (6.96 X 10(8) cfu/mL) was constructed for selecting high activity of anti-CrylF toxin single-chain antibody (a single-chain variable fragment, scFv) by biopanning. A total of 16 positive monoclonal phage scFv's were obtained after 4 rounds of panning, which were identified by enzyme-linked immunosorbent assay (ELISA), polymerized chain reaction, and DNA sequencing. The most positive phage scFv (named RF4) was expressed in Escherichia coli HB2151, and a soluble protein of approximately 30 kDa was purified with sodium dodecyl sulfate polyacrylamide gel electrophoresis. An indirect competitive ELISA (IC-ELISA) was established on the basis of purified soluble RF4-scFv for CrylF toxin. It indicated the 50% inhibition of the control (IC50) was 11.56 ng/mL and the detection limit (IC10) was 0.18 ng/mL and showed weak cross-reactivities for CrylAb (2.8%), CrylAc (1.3%), and Cry1B, Cry1C, Crylle, and Cry2A (less than 0.1%). It was found that IC-ELISA detected Cry1F toxin spiked in rice, wheat, corn, and soil samples with good accuracy, stability, and repeatability. The recoveries were in the range of 80.2-99.6%, and the coefficients of variation were in the range of 2.5-10.0%. These results showed that IC-ELISA based on scFv from the immunized rabbit phage display library was promising for specific detection of Cry1F toxin in agroproducts and environmental samples.

分类号： R15`S

  • 相关文献

[1]Development of Bt Rice and Bt Maize in China and Their Efficacy in Target Pest Control. Liu, Qingsong,Peng, Yufa,Li, Yunhe,Hallerman, Eric. 2016

[2]Different toxicity of the novel Bacillus thuringiensis (Bacillales: Bacillaceae) strain LLP29 against Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae). Zhang, Lingling,Tang, Baozhen,Huang, Zhipeng,Liu, Zhaoxia,Huang, Tianpei,Guan, Xiong,Huang, Enjiong,Gelbic, Ivan,Xu, Lei.

[3]Construction of scFv phage display library with hapten-specific repertories and characterization of anti-ivermectin fragment isolated from the library. Zhang, Xiao,Zhang, Cunzheng,Liu, Yuan,Yu, Xiangyang,Liu, Xianjin. 2010

[4]Establishment of a sensitive time-resolved fluoroimmunoassay for detection of Bacillus thuringiensis Cry1Ie toxin based nanobody from a phage display library. Xu, Chongxin,Zhang, Cunzheng,Zhang, Xiao,Zhong, Jianfeng,Liu, Yuan,Hu, Xiaodan,Lin, Manman,Liu, Xianjin,Liu, Xiaoqin. 2017

[5]Polyclonal antibodies to grain gliadins and their application in wheat quality prediction. Li, Q. Y.,Song, X. Y.,Zhang, E. Y.,Pei, Y. H.,Li, Q. Y.,Ji, K. M.,Yan, Y. M.. 2008

[6]Quantification of toxins in a Cry1Ac+CpTI cotton cultivar and its potential effects on the honey bee Apis mellifera L.. Peng Han,Chang-Ying Niu,Chao-Liang Lei,Jin-Jie Cui,Nicolas Desneux. 2010

[7]Serological Survey of Canine Leptospirosis in Southern China. Liu, Mingchao,Liu, Jianzhu,Wang, Lin,Wang, Zhenyong,Wang, Shujing,Yang, Dubao,Chai, Tongjie,Shi, Dayou,Guo, Shining,Liao, Shenquan,Sun, Mingfei. 2012

[8]A monoclonal antibody-blocking enzyme-linked immunosorbent assay for the detection of serovar-specific antibodies to Haemophilus paragallinarum. Zhang, P,Blackall, PJ,Yamaguchi, T,Iritani, Y. 1999

[9]Tier-1 assays for assessing the toxicity of insecticidal proteins produced by genetically engineered plants to non-target arthropods. Romeis, Joerg,Wu, Kong-Ming,Peng, Yu-Fa,Romeis, Joerg. 2014

[10]Bioaccumulation of microcystin and its oxidative stress in the apple (Malus pumila). Chen, Jianzhong,Zhang, Haiyang,Wang, Chenyi,Zhou, Guoqing,Han, Zhiping,Chen, Jianzhong,Liu, Zhili,Dai, Jian. 2010

[11]Development and evaluation of a VP1-ELISA for detection of antibodies to duck hepatitis type 1 virus. Zhang, Tingting,Zhang, Yun,Meng, Fanyi,Li, Xiaojun,Kong, Xiangang,Liu, Ming,Hou, Zhenzhong,Feng, Xinchang. 2010

[12]Seroprevalence and Risk Factors of Fascioliasis in Yaks, Bos grunniens, from Three Counties of Gansu Province, China. Zhang, Xiao-Xuan,Ma, Jian-Gang,Zheng, Wen-Bin,Zhao, Quan,Zhang, Xiao-Xuan,Feng, Sheng-Yong,Ma, Jian-Gang,Zheng, Wen-Bin,Yin, Ming-Yang,Qin, Si-Yuan,Zhou, Dong-Hui,Zhu, Xing-Quan. 2017

[13]Development of a Synthetic Peptide ELISA Assay for the Detection of Foot-and-Mouth Disease Virus Nonstructural Protein Antibodies. Yang Ji-fei,Yang Su-zhen,Yang Yan-yan,Zhi Ai-min,Zhao Dong,Zhi Yu-bao,Xing Guang-xu,Deng Rui-guang,Chai Shu-jun,Zhang Gai-ping,Yang Su-zhen. 2010

[14]Preliminary serological evidence for Schmallenberg virus infection in China. Zhai, Shao-Lun,Lv, Dian-Hong,Wen, Xiao-Hui,Zhu, Xue-Liang,Yang, Yan-Qiu,Chen, Qin-Ling,Wei, Wen-Kang. 2018

[15]Seroprevalence of bovine theileriosis in northern China. Li, Yaqiong,Liu, Zhijie,Liu, Junlong,Yang, Jifei,Li, Qian,Guo, Pengfei,Guan, Guiquan,Liu, Guangyuan,Luo, Jianxun,Yin, Hong,Li, Youquan,Yin, Hong. 2016

[16]Identification of Theileria uilenbergi immunodominant protein for development of an indirect ELISA for diagnosis of ovine theileriosis. Liu, Zhijie,Kullmann, Birgit,Abdo, Jassim,Salih, Dialeldin,Ahmed, Jabbar,Seitzer, Ulrike,Liu, Zhijie,Wang, Zijian,Yin, Hong,Luo, Jianxun,Zhang, Bao. 2010

[17]Development of a Competitive Indirect Enzyme-Linked Immunosorbent Assay Based on Monoclonal Antibodies for the Detection of 2-Dodecylcyclobutanone in Irradiated Beef. Zhao, Yueliang,Wang, Feng,Li, Weiming,Ha, Yiming. 2013

[18]Development of an enzyme-linked immunosorbent assay for podophyllotoxin. Xu, DM,Yu, XY,Liu, YQ,Feng, JT,Pan, LG,Liu, XJ,He, J,Zhang, X. 2005

[19]Enzyme-Linked Immunosorbent-Assay for Deoxynivalenol (DON). Ji, Fang,Li, Hua,Xu, Jianhong,Shi, Jianrong. 2011

[20]Analysis of the relationship between codon frequency of prolactin gene and laying performance in five chicken breeds. Liu Hua-Gui,Wang Xiao-Hua,Liu Yu-Fang,Zhao Xing-Bo,Li Ning,Wu Chang-Xin. 2007

作者其他论文 更多>>

微信小程序